NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL
Year:2016 | Volume:6 | Issue:21|Papers Count:17

One of the common causes of cancer-related mortalities worldwide is lung cancer and more than 1 million people annually die from it. Lung cancers are divided into Non-small cell lung cancer (NSCLC) and small cell lung carcinomas (SCLCs) which are different in several characteristics like biological behavior, response to therapy and genetic alterations.Resistance to conventional chemotherapy, especially in NSCLC patients is a major challenge in the treatment process. Therefore, development of novel therapeutic strategies is essential for improvement of survival rate. Combination of different therapeutic methods is an approach which can maximize efficiency of the treatment.Small interfering RNA (siRNA) has been reported to have a huge potential for the treatment or prevention of various lung diseases. Once the RNA molecules have successfully entered the target cells, they could inhibit the expression of specific gene sequence through RNA interference (RNAi) mechanism and generate therapeutic effects. The biggest obstacle to translating siRNA therapy from the laboratories into the clinics is delivery. An ideal delivery system should protect the siRNA from enzymatic degradation, facilitate cellular uptake and promote endosomal escape inside the cells, with negligible toxicity.Very limited work has been done on the formulation of siRNA for inhalation which is believed to be the direction for future development. This review article introduces lung cancer and siRNA as a new treatment, and also studies the challenges of siRNA delivery to lung cells by non-viral carriers. Furthermore, it surveyed some nano carriers for pulmonary siRNA delivery with lipid, polymer and peptide basic.

Aim and Background: HIV protease plays an important role in the maturation of the virus and host immune response stimulation; therefore it is important as a therapeutic target and can be used in diagnostic tests. In previous studies, producing rPR has been faced with problems such as toxicity, hydrophobic and purification. The purpose of this study is the use of expression system pET102/D. TOPO to overcome the mentioned problems.Materials and Methods: Protease gene was isolated from the serum of an infected individual. Cloning process was performed by TOPO Cloning system. Protease gene transformed into E.coli BL21and induced with IPTG. Produced recombinant protein was purified by affinity chromatography (Ni-NTA column). Protein concentration was checked by BCA protein assay kit. Recombinant protein was identified by SDS-PAGE and western-blotting.Results: HIV-1 protease gene Cloning using TOPO Cloning vector expression system pET102 / D was confirmed by PCR and sequencing. rPR concentration after purification was 60 to 85 micrograms per milliliter. rPR Expression and Immunogenicity by SDS-PAGE and Western blotting were confirmed.Conclusion: Expression and production of rPR in soluble form was a success. In used Cloning system, because of the expression of rPR in fused form with thioredoxin and histidine tags, problems of toxicity, hydrophobicity and purification were noticeably solved. According to the generated rPR, it can be used to evaluate the diagnostic tests and for designing protease inhibitors in subsequent studies.

Aim and Background: whole-mount in situ hybridization (wmISH) is a powerful tool to visualize the gene transcript in embryonic tissue. This study aimed at setting up wmISH using digoxygenine-labled RALDH2 antisense riboprobe.Methods & Materials: The early developing chick embryos were fixed and dehydrated upon dissecting. Following bleaching and protein kinase K treatment, the embryos were hybridized with hybridization buffer and then with the buffer containing RALDH2 antisense riboprobe for 36h. Upon washing with SSC solution and incubation with anti-digoxygenin Ab, the embryos were treated with NBT/BCIP and transferred to NTMT buffer. Some embryos received sense ribobrobe and negative control was considered as a group not treated with any riboprobes.Results: Following binding the antisense riboprobe to RALDH2 transcript, blue color appeared in somites and neural tube as well as the cardiac tube. No color was detected in the embryonic tissue treated with the sense riboprobe or in untreated group.Conclusion: Using wmISH, we can precisely detect location and rate of the gene transcription in the developing embryo.

Aim and Background: Hepatocellular carcinoma (HCC) -an aggressive malignancy with high rate of mortality- is the third cause of cancer related death, worldwide. Cisplatin is a chemotherapy drug used for treatment of HCC. However, serious side effects of drug limit the therapeutic administer dose. Nanotechnology based devices such as liposome nanoparticles are capable to overcome these drawbacks and simultaneously improve the efficacy of drugs.Materials and Methods: Cisplatin encapsulated liposomal nanoparticles were successfully constructed using lecithin, Cholesterol, PEG3350and CisPlatin. After characterization in vitro efficacy of nanodrug was evaluated with IC50 Pharm Microsoft.Results: The size and zeta potential of nanoparticles were determined to be 470 nm and -20 mV.The cytotoxicity of drug loaded nanoparticles was increased by 33% compared to the free drug on the Hepatocellular carcinoma cell line HepG2 using MTT assay.Conclusion: The results of study suggested evaluating the efficacy of nanodrug on the animal model of HCC and also the cytotoxicity of pegilated nanoliposome drug is more than the free drug.

Aim and Background: In order to evaluate the effect of priming on seed germination Burley 21 tobacco varieties, a research was conducted in the autumn of 1392 in Urmia Tobacco Research Center.Material and methods: In the first part, a completely randomized design with 26 treatments and 3 replications were conducted. Treatments were one control and 4 levels of polyethylene glycol (0.5, 1, 1.5 and 2%) at different periods of 1, 2, 3, 5 and 10 days, and water (priming) at different periods of 1, 2, 3, 5 and 10 days. In the second part, a factorial completely randomized design was conducted with 3 factors and 3 replications for the first factor, the concentrations of -0.5, -1, -1/5 and-2 MPa polyethylene glycol and distilled water, and for the second factor they were 1, 2, 3, 5 and 10 days priming and salinity; for the third factor were 1, 2, 3 and 4 dS m for per was petri dish.Results: In the first part of experiment, the treatments on time and germination percentage at 1% level had a significant effect. The mean time of germination using tobacco seed priming with polyethylene glycol and distilled water significantly decreased. Germination in the priming effect for 3, 5 and 10 days significantly was reduced. In the second part of the experiment, seed priming had significant effect on germination time, germination rate, germination index, germination rate and germination percentage at 1 % level, time duration of seed priming on germination time, germination rate had significant effect at 1% level. Germination time in petri dish significantly increased with increasing salinity; while the germination rate and speed germination index declined. In conclusion, priming tobacco, resulting in improved germination rate, rate of germination and speed germination. The best priming time was 1 and 1.5 % polyethylene glycol and the duration of a day.Conclusions: Considering the importance of tobacco seed germination rate, it is required to use concentrations of 1 or 1.5% of polyethylene glycol to be treated one day before.

Aim and Background: Rachycentron Canadum is one of the most important commercial species in the Persian Gulf. This study focuses on molecular investigation of mentioned species in order to find and introduce the genetic differentiations and also probable genotypes for monitoring and managing the genetic resources of populations in three major catch areas in the Persian Gulf and the Oman Sea.Materials and methods: 120 individuals of this species were caught from the northern part of the Persian Gulf and Oman Sea. DNA extraction was performed from the dorsal part of the fin of 54 Individuals using Phenol-Chloroform method. Polymerase chain Reaction (PCR) was performed using a set of primers which designed based on NADH dehydrogenase (ND2) gene.Results: The haplotype diversity from the populations of Bushehr 0. ± 0, Sistan Baluchestan 0. ± 0.3 and mean diversity was recorded 0. ± 0.015. Also haplotype diversity from the populations of Khuzestan and Hormozgan were 0. ± 0.04 and 0. ± 0.04 respectively and the mean diversity was recorded 0. ± 0.04. haplotype diversity from the populations of Khuzestan and Bushehr were 0. ± 0.04 and 0. ± 0 respectively and the mean diversity was recorded 0. ± 0.02.Discussion: The study showed that some haplotypes of the populations belonging to Khuzestan were setting in one clade with common ancestor compared with the other population (Bushehr, Hormozgan and Sistan population) shown in another clade indicating that restricted genetic flow between Khuzestan and other studied areas. This study confirmed that several factors including barriers (mangrove forests) and marine currents can have an effect on the population diversity of this species in the Persian Gulf and Oman Sea.

Aim and Background: Mentha longifolia with the generic name "Wild Mint, is one of the oldest medicinal plants which have been widely used in Iranian food and medicine. This study was performed to extract the essential oil of Mentha longifolia and analysis of its components and also antioxidant and antibacterial properties of its oil and non-polar compounds, extracted by n-hexan.Materials and Methods: In this study, the essential oil of M. longifolia was extracted using SDE technique for the first time. The obtained essential oil was analyzed using GC-Mass and its components are identified. Antioxidant activity of the oily compounds was measured via DPPH free radical scavenging activity; while, antibacterial effects were evaluated using disc diffusion method by measuring inhibition zone diameter evaluating n-hexane extract.Results: The major components of the oil are d-carvone (57.2%) and limonene (15.7%). Some of the essential oil components, identified in this study, are not similar to those mentioned in the previous reports. The results indicate that despite non-polar nature of extract solvent, n-hexane extract has a noticeable inhibitory power on DPPH free radical. Also, it shows very high antimicrobial properties on gram-positive and gram-negative bacteria, which are comparable with standard antibiotics.Conclusions: This study shows that components extracted by hexane as solvent, despite oily nature of compounds with low polarity, have antioxidant activity and also significant anti-microbial potential which have not been reported for M. longifolia in the literature.

Introduction: Nowadays, using nanocarriers and targeted drug delivery have attracted more attention. Nanocarriers, especially gold nanoparticles have been presented as an appropriate choice for targeted drug delivery. Today, applying biological systems like microorganisms in synthesizing gold nanoparticles has been the focus of attention. Biosynthesis and aggregation of nanoparticles using clean biological systems which are biocompatible and non-toxic are known as green methods, especially compared with other chemical methods of synthesizing nanoparticles.Materials and Methods: In this study, Staphylococcus aureus was used for the biosynthesis of gold nanoparticles and the effects of their antimicrobial and cytotoxicity were examined.Results: Perfectly spherical gold nanoparticles with an average size of 25 nm were synthesized. Techniques using different concentrations of gold nanoparticles were studied by MTT assay, and the concentration of more than half of the cells survived. MIC and MBC gold nanoparticles microbial tests for vancomycin-resistant Enterococcus faecium utilized for all concentrations of nanoparticles were dead bacteria.Conclusion: Using Gold Nanoparticles omitted vancomycin resistance in vancomycin-resistant Enterococci.

Aim and background: Traditionally Nocardia asteroides was considered the predominant species of causing nocardiosis. The improved identification of isolates using molecular techniques have shown that the genus exhibits considerable taxonomic complexity and phenotypic base identification can be ambiguous. The aim of this study was to assess the species distribution of Nocardia strains mostly recovered from patients suspected of having tuberculosis, during three years period (2009- 2012).Material and methods: The clinical isolates were identified to species level using conventional tests and genotypic methods using single and multi-locus sequence analysis (MLSA) of 16S rRNA, gyrB and secA1 genes. Nocardiosis was diagnosed in 46 patients.Results: The most frequent underlying condition were organ transplantation (6 patients, 13%), cancer (6 patients, 13%), human immunodeficiency virus (HIV) (6 patients, 13%), non-infectious chronic lung disease (5 patients, 10.8%) and tuberculosis (4 patients, 8.7%). Nocardia species was recovered from 46 different clinical specimens, the most common of which was bronchoalveolar lavage (BAL) (43.5%). Eleven different Nocardia species were identified: N. asteroides (n=12), N. cyriacigeorgica (n=9), N. farcinica (n=7), N. wallacei (n=6), N. carnea (n=3), N. otitidiscaviarum (n=3), N. abscessus (n=1), N. arthritidis (n=1), N. kruczakiae (n=1), N. nova (n=2) and N. veterana (n=1).Conclusion: In conclusion, infection caused by Nocardia species appears to be more common than is generally appreciated. The current study provides further evidence that Nocardia species are capable of causing a wide range of human diseases in healthy and immunocompromised patients. MLSA is a reliable method for accurate species identification of Nocardia isolates and would be more feasible for routine use in clinical laboratories.

Aim and Background: Given the importance of the intertidal benthic communities as well as increasing urban and industrial facilities on the coast, this study was carried out to obtain basic information required in the control of the coast, environmental.Material and Methods: Sampling was conducted in Bandar e Abbas, Bandar e Jask and Bandar e Lengeh by a powerboat in 2011 seasonally.Conclusion: In this study Polycheata, Oligocheata, Crastacea, Mollusca, Nematoda, Nemerteana, Foraminiferida, Ophiuroidea and others were carried out. Results showed that Crustacea had the maximum density with amount of 333950 indm-2.Discussion: Any changes to coastal ecosystems and intertidal zone will result in a change in the factors that govern the benthic communities as indicators of environmental changes as well as the dynamics of the visible world, evaluating and calculating.Results: Due to Macrobenthos density and biodiversity indicators, it seems that a significant change in the density and biodiversity after algal bloom did not exist therefore; it had been before on the coast of Hormozgan.

Background and Objectives: The immune antigen of Bacillus anthracis is a protein that can attach to the surface receptor of all human cells. At the surface of cancer cells there is a receptor that activates the uPA (Urokinase plasminogen) that do not exist in normal human cells. This research was conducted to change the site of attachment of PA gene with making direct mutation so that it can attach only to the cancer cells.Material and Methods: In this study, pMNA1 plasmid containing PA gene was extracted from its host by basic lyse. Subsequently, phosphorylized primers and Overlap Extention PCR were used to make mutation on PA gene. The mutated segment was directly cloned in PTZ57R carrier and transferred to DH5α strain of E. coli. By catalyst enzymes KpnI and SalI the mutated PA gene was extracted and transferred to pWB980and by electroporation method it was transferred to WB600 strain.Results: In this study mutation was occurred in sequences of PA gene by SOE PCR method resultingin a change in the genetic code of amino acid 194. The occurrence of mutation was confirmed by determining of base sequences.Conductions: Cancer is a serious disease that in many cases leads to death. One of the treatment methods of cancer is bacterial toxins if only cancer cells receive them. Therefore, using changed PA can be a hope in cancer treatment as it attaches only to cancer cells.

Aim and background: The aim of this multicenter study was to determine the antibiotic susceptibility of clinically isolated Nocardia species.Material and methods: One hundred twenty-seven patients with nocardiosis were randomly selected from 5 provinces of Iran. Molecular diagnosis of Nocardia species was performed using 16S rRNA sequencing. Antimicrobial susceptibility testing was performed following the Clinical and Laboratory Standards Institute recommendations.Results: Thirty-five N. cyriacigeorgica, 30 N. asteroides, 26 N. farcinica, 12 N. otitidiscaviarum, and 10 N. abscessus cultures were studied. All isolates were susceptible to linezolid. All isolates of N. cyriacigeorgica, N. asteroides, N. abscessus, and N. otitidiscaviarum were susceptible to trimethoprim-sulfamethoxazole, while 8% of N. farcinica isolates were resistant to this drug. All N. otitidiscaviarum isolates were highly resistant to imipenem, but N. cyriacigeorgica, N. asteroides, N. farcinica, and N. abscessus were only moderate resistant.Conclusion: The susceptibility patterns vary with different species of Nocardia. Resistance to trimethoprim-sulfamethoxazole in Iran is low and this drug should be first line therapy, unless drug susceptibility testing shows resistance. Linezolid also covers Nocardia well and could be a second line agent.

Aim and Background: Vanadium compounds have both inhibitory and antitumor effects against chemical agents that cause cancer in a variety of cell lines. Also, previous studies demonstrated that curcumin in low concentrations, induce apoptosis and inhibit the cancer cells proliferation. The aim of this study is the synthesis of new complex of vanadium-curcumin and evaluation of anti-proliferative and apoptotic effects of this compound.Material and methods: In this study, we synthesized the novel complex of vanadium, (1, 7-bis [4-hydroxy-3-methoxyphenyl] -1, 6-heptadiene-3, 5- dionato) acetylacetonato oxovanadium (IV). The cytotoxicity of this complex was screened for antitumor activity against human breast cancer cells (MCF-7) human cervix epithelial carcinoma (HeLa), human colon cancer cell line (HT-29), human leukemia cell line (K-562), mouse neuroblastoma cell line (Neuro-2a) and normal mouse fibroblast cell line (L929) and using cisplatin as a comparative standard by MTT assay and Flow cytometry. The effect of synthesized compound on mouse fibroblast cell line (L929) was evaluated as control, simultaneously.Results: The IC50 ± SD (mM) values, calculated for synthesized complex against HeLa, MCF-7, HT-29, K-562, Neuro-2a and L929 cell lines are 5.65 ± 1.23, 2.48 ± 1.05, 42.6 ± 5.19, 3.99 ± 1.12, 65.3 ± 6.89 and 106.7 ± 8.98 respectively, while The IC50 ± SD (mM) values, calculated for cisplatin as a comparative standard drug in chemotherapy against these cell lines, are 0.45 ± 0.12, 6.50 ± 2.11, 15.4 ± 4.93, 24.0 ± 2.93 and>200, respectively. Also the flow cytometry results for synthesized complex showed a high population of apoptotic cell (58.46%) and nearly 1.2-fold higher than cisplatin (48.03%) at the same concentration.Conclusion: Our results showed that investigated complex exhibited higher cytotoxic activities against human breast cancer cells (MCF-7) and human leukemia cell line (K-562) compared to other cell lines. Our results confirmed that the synthesized compound have no cytotoxicity effects on mouse fibroblast cell line (L929). Also the flow cytometry results confirm that vanadium complex induce apoptosis in human leukemia cell line (K-562).

Aim and Background: Chemotherapy is a category of cancer treatment that uses chemical agents. The harmfulness of chemotherapy on normal cells by killing cells is the most common side-effects of chemotherapy. Characterization and selection of new powerful chemotherapy agents which have low side effects are important. Indoloquinazoline has a variety of biological activities such as antimicrobial activities and cytotoxic properties. In this study, cytotoxic effect of synthetic indoloquinazoline compound was studied on breast cancer cell line and compared with cisplatin.Materials and Methods: Anti tumor activity of indoloquinazoline compound and cisplatin was studied on breast cancer cell line, MCF-7, by MTT assay.Results: Indoloquinazoline compound showed antitumor activity with IC50 of 6.83 mg/ml by being Compared with 39.13 mg/ml of cisplatin, as a kind of standard drug on MCF-7 cell line.Conclusion: The results showed indoloquinazoline is cytotoxic more than cisplatin on MCF-7 cell line.